Dual selection of Beauveria bassiana strains and complex substrate media for the massive production of submerged propagules with activity against the eucalyptus bronze bug Thaumastocoris peregrinus

The bronze bug Thaumastocoris peregrinus is an invasive pest, affecting Eucalyptus plantations worldwide. Although its natural enemy Cleruchoides noackae has been tested for the biological control of this pest, other strategies like the use of native entomopathogenic fungi are needed. For this, native virulent fungal isolates should be selected, massively multiplied in an efficient way, and prepared to obtain a stable product. Isolates of native Beauveria bassiana obtained from T. peregrinus and from different collections were screened for their virulence towards this insect and for their amenability to be massively produced in a low-input liquid submerged fermentation and prepared as a dry powder. Three out of six virulent strains were suitable for their massive production in a 2% corn flour suspension, achieving 10⁹ submerged propagules/g of dehydrated preparation. The LC50 achieved by the dry submerged propagules did not differ from the LC50 of fresh aerial conidia. The proposed dual selection of strain and a complex substrate, and the procedures leading to the production of a dry preparation, allowed for high viability and virulence of the fungal spores of three strains.     

Soil and entomopathogenic fungi with potential for biodegradation of insecticides: degradation of flubendiamide in vivo by fungi and in vitro by laccase

Purpose

Flubendiamide is a highly toxic and persistent insecticide that causes loss of insect muscle functions leading to paralysis and death. The objective was to screen for filamentous fungi in soils where insecticides had been applied, to isolate entomopathogenic fungi from insect larva (Anticarsia gemmatalis) that infest soybean crops, and to use these in biodegradation of insecticides.

Method

Filamentous fungi were isolated from soils, and growth inhibition was evaluated on solid medium containing commercial insecticides, Belt® (flubendiamide) and Actara® (thiamethoxam). A total of 133 fungi were isolated from soil and 80 entomopathogenic fungi from insect larva. Based on growth inhibition tests, ten soil fungi, 2 entomopathogenic fungi, and Botryosphaeria rhodina MAMB-05 (reference standard) were selected for growth on commercial insecticides in solid media. Fungi were grown in submerged fermentation on media containing commercial insecticides and assayed for laccase activity.

Result

Isolates JUSOLCL039 (soil), JUANT070 (insect), and MAMB-05 performed best, and were respectively inhibited by 48.41%, 75.97%, and 79.23% when cultivated on 35 g/L Actara®, and 0.0, 5.42%, and 43.39% on 39.04 g/L Belt®. JUSOLCL039 and JUANT070 were molecularly identified as Trichoderma koningiopsis and Neurospora sp., respectively. The three fungal isolates produced laccase constitutively, albeit at low activities. Fungal growth on pure flubendiamide and thiamethoxam resulted in only thiamethoxam inducing high laccase titers (10.16 U/mL) by JUANT070. Neurospora sp. and B. rhodina degraded flubendiamide by 27.4% and 9.5% in vivo, while a crude laccase from B. rhodina degraded flubendiamide by 20.2% in vitro.

Conclusion

This is the first report of fungi capable of degrading flubendiamide, which have applications in bioremediation.

     

Molecular characterization,virulence and horizontal transmission of Beauveria pseudobassiana from Dendroctonus micans (Kug.) (Coleoptera: Curculionidae)

The great spruce bark beetle, Dendroctonus micans (Kugelann) (Coleoptera: Curculionidae), has been a potential threat for Turkey and the entire Eurasian spruce forests for many years. Control strategies which have been applied so far are still insufficient to prevent its damage. A previous study has shown that a Beauveria isolate (ARSEF 9271) proved to be an efficient microbial control agent against the great spruce bark beetle. In this study, this isolate was identified as B. pseudobassiana based on the partial sequence of EF1‐α and ITS sequence. A conidial suspension (1 × 10⁸/ml) of this fungus caused 100% mortality on both larvae and adults of D. micans within 5 and 6 days, respectively. Also, it caused 100% mycosis value on both larvae and adults. Mortality values of horizontal transmission experiments between larvae and adults which were contaminated with 1 × 10⁶/ml spore suspension at 25%, 50%, 75% and 100% rates were determined as 100% after 15 days at 20°C under the laboratory conditions. We also determined the decrease of the damage in spruce wood block (15 × 25 cm) when the contamination rate of the larvae was increased. Our results indicate that B. pseudobassiana ARSEF 9271 seems to be a very promising biocontrol agent against D. micans.     

DNA bar-coding reveals source and patterns of <Emphasis Type="Italic">Thaumastocoris peregrinus</Emphasis> invasions in South Africa and South America

Thaumastocoris peregrinus is a recently introduced invertebrate pest of non-native Eucalyptus plantations in the Southern Hemisphere. It was first reported from South Africa in 2003 and in Argentina in 2005. Since then, populations have grown explosively and it has attained an almost ubiquitous distribution over several regions in South Africa on 26 Eucalyptus species. Here we address three key questions regarding this invasion, namely whether only one species has been introduced, whether there were single or multiple introductions into South Africa and South America and what the source of the introduction might have been. To answer these questions, bar-coding using mitochondrial DNA (COI) sequence diversity was used to characterise the populations of this insect from Australia, Argentina, Brazil, South Africa and Uruguay. Analyses revealed three cryptic species in Australia, of which only T. peregrinus is represented in South Africa and South America. Thaumastocoris peregrinus populations contained eight haplotypes, with a pairwise nucleotide distance of 0.2–0.9% from seventeen locations in Australia. Three of these haplotypes are shared with populations in South America and South Africa, but the latter regions do not share haplotypes. These data, together with the current distribution of the haplotypes and the known direction of original spread in these regions, suggest that at least three distinct introductions of the insect occurred in South Africa and South America before 2005. The two most common haplotypes in Sydney, one of which was also found in Brisbane, are shared with the non-native regions. Sydney populations of T. peregrinus, which have regularly reached outbreak levels in recent years, might thus have served as source of these three distinct introductions into other regions of the Southern Hemisphere.     

Characterization and virulence of entomopathogenic fungi from sunn pests in Turkey

The sunn pests (Eurygaster spp.) which are important pests of wheat and barley, can lead to 100% yield loss in the case of an epidemic. This study was conducted to characterize the entomopathogenic fungi, obtained from the sunn pests collected from different overwintering areas of Turkey and determine their pathogenicity. Seventeen Beauveria pseudobassiana, 12 Beauveria bassiana, 16 Cordyceps farinosa, 1 Purpureocillium lilacinum, and 2 Clonostachys rosea species were identified. Pathogenicity tests were performed with three B. pseudobassiana, two B. bassiana, four C. farinosa, and one P. lilacinum isolates. B. pseudobassiana isolates BB34 and AF4 caused 100% death on the 9th day, and another B. pseudobassiana isolate E512 caused 100% death on the 12th day. C. farinosa isolate KOKA2 caused 95.8% death on the 15th day. Ultimately, B. pseudobassiana was determined to be the most prevalent and virulent species on sunn pests.     

Identification and Pathogenicity of Lasiodiplodia Species from Eucalyptus urophylla × grandis,Polyscias balfouriana and Bougainvillea spectabilis in Southern China

Species of Lasiodiplodia are important pathogens of a wide variety of plants covering a wide geographical distribution. These fungi can be associated with different symptoms such as stem cankers, shoot blights, fruit rots, dieback and gummosis. Diseases caused by Lasiodiplodia were surveyed on Eucalyptus urophylla × grandis, Polyscias balfouriana and Bougainvillea spectabilis in a nursery in southern China. Based on morphology characteristics and phylogenetic analyses of ITS rDNA sequences and translation elongation factor 1‐alpha (TEF‐1α) gene regions, four species of Lasiodiplodia were identified. Lasiodiplodia theobromae was identified from E. urophylla × grandis, P. balfouriana and B. spectabilis. L. hormozganensis, L. iraniensis and L. pseudotheobromae were identified from B. spectabilis. To our knowledge, with the exception of L. theobromae on E. urophylla × grandis, this study represents the first report of these fungi on the host plants. Pathogenicity tests showed that all Lasiodiplodia spp. obtained in this study are virulent to E. urophylla × grandis and B. spectabilis, and L. theobromae was virulent to P. balfouriana.     

Lethal activity of beauvericin,a Beauveria bassiana mycotoxin,against the two‐spotted spider mites,Tetranychus urticae Koch

The entomopathogenic fungi Beauveria bassiana (Balsamo) Vuillemin is known to produce a broad range of secondary metabolites. Beauvericin, a cyclic hexadepsipeptide, is the best known mycotoxin produced by B. bassiana; however, reports discussing the insecticidal activity of beauvericin per se are limited. In this study, we assessed the lethal activity of beauvericin against Tetranychus urticae Koch (Acarina: Tetranychidae). In addition, screening for suitable application of the mycotoxin against T. urticae on greenhouse strawberries is discussed. Beauvericin was able to control successfully T. urticae where concentrations of 10, 100 and 1,000 µg/g recorded mortalities of 84%, 100% and 100%, respectively, against motile stages. Furthermore, beauvericin inhibited egg hatching up to 83.3%, 69.3% and 53.3%, respectively, using the same concentrations under laboratory conditions. Under greenhouse conditions, the efficacy recorded was 52.6%, 85.7%, 72.4% and 72.4% at 1, 3, 7 and 10 days post‐inoculation, respectively. Beauvericin was efficacious under greenhouse conditions since the application increased strawberry yields while showing no phytotoxicity and ecotoxicological risk. Resistance to beauvericin was not detected initially at the unselected strain of T. urticae. Yet, the laboratory selection of populations of T. urticae exposed to beauvericin resulted in relatively resistant T. urticae strain that displayed no cross‐resistance to cyflumetofen and bifenazate. The acaricidal activity of beauvericin documented in this study would increase the efficacy of integrated pest management strategies.     

Virulence of entomopathogenic nematodes and their symbiotic bacteria against the hazelnut weevil Curculio nucum

The virulence of different entomopathogenic nematode strains of the families Steinernematidae and Heterorhabditidae, isolates from Catalonia (NE Iberian Peninsula), and their symbiotic bacteria was assessed with regard to the larvae and adults of the hazelnut weevil, Curculio nucum L. (Coleoptera: Curculionidae). The nematode strains screened included one Steinernema affine, five Steinernema feltiae, one Steinernema carpocapsae, one Steinernema sp. (a new species not yet described) and one Heterorhabditis bacteriophora. The pathogenicity of all the strains of nematodes was tested on larvae and only four of them on adults of the hazelnut weevil. Larval mortality ranged from 10% with S. affine to 79% with Steinernema sp. Adult mortality was higher in S. carpocapsae, achieving 100% adult weevil mortality. The pathogenicity of the symbiotic bacteria Xenorhabdus bovienii, X. kozodoii, X. nematophila and Photorhabdus luminescens was studied in larvae and adults of C. nucum. In the larvae, X. kozodoii showed a LT50 of 22.7 h, and in the adults, it was 20.5 h. All nematodes species except S. affine tested against larvae showed great potential to control the insect, whereas S. carpocapsae was the most effective for controlling adults.     

A genome scan of diversifying selection in Ophiocordyceps zombie‐ant fungi suggests a role for enterotoxins in co‐evolution and host specificity

Identification of the genes underlying adaptation sheds light on the biological functions targeted by natural selection. Searches for footprints of positive selection, in the form of rapid amino acid substitutions, and the identification of species‐specific genes have proved to be powerful approaches to identifying the genes involved in host specialization in plant‐pathogenic fungi. We used an evolutionary comparative genomic approach to identify genes underlying host adaptation in the ant‐infecting genus Ophiocordyceps, which manipulates ant behaviour. A comparison of the predicted genes in the genomes of species from three species complexes—O. unilateralis, O. australis and O. subramanianii—revealed an enrichment in pathogenesis‐associated functions, including heat‐labile enterotoxins, among species‐specific genes. Furthermore, these genes were overrepresented among those displaying significant footprints of positive selection. Other categories of genes suspected to be important for virulence and pathogenicity in entomopathogenic fungi (e.g., chitinases, lipases, proteases, core secondary metabolism genes) were much less represented, although a few candidate genes were found to evolve under positive selection. An analysis including orthologs from other entomopathogenic fungi in a broader context showed that positive selection on enterotoxins was specific to the ant‐infecting genus Ophiocordyceps. Together with previous studies reporting the overexpression of an enterotoxin during behavioural manipulation in diseased ants, our findings suggest that heat‐labile enterotoxins are important effectors in host adaptation and co‐evolution in the Ophiocordyceps entomopathogenic fungi.     

Efficacy of commercially available predators,nematodes and fungal entomopathogens for augmentative control of Drosophila suzukii

The recent arrival of Drosophila suzukii, an invasive pest of soft‐skinned fruit with a wide host range, has resulted in increased production costs for growers and the need for additional insecticide applications each growing season. There are few effective organic insecticides for D. suzukii, and insecticide use in conventional farms may be disruptive to natural enemies, suggesting a need for effective biological control to combat D. suzukii. Commercially available natural enemies were evaluated for their potential use in augmentative releases, including: the predators Orius insidiosus and Dalotia coriaria; the entomopathogenic fungi Metarhizium anisopliae, Beauveria bassiana and Paecilomyces fumosoroseus; and the entomopathogenic nematodes Heterorhabditis bacteriophora, Steinernema feltiae and S. carpocapsae. This suite of natural enemies was chosen to target D. suzukii adults as well as larvae in hanging or dropped fruit. Of the cultured fungal strains tested, only M. anisopliae significantly decreased D. suzukii survival, but it had low residual activity and no effect on D. suzukii fecundity. O. insidiosus decreased D. suzukii survival in simple laboratory arenas but not on potted blueberries or bagged blueberry branches outdoors. D. coriaria did not decrease D. suzukii survival in infested blueberries in simple laboratory arenas. The nematodes tested showed low infection rates and were not able to affect D. suzukii survival. Although this suite of natural enemies showed limited ability to suppress D. suzukii under the tested conditions, these and related natural enemies are present as part of the endemic natural enemy community in agricultural fields, where they may contribute to D. suzukii suppression.     

Quantification of Alternaria,Cladosporium, Fusarium and Penicillium verrucosum in Conventional and Organic Grains by qPCR

We analysed the levels of Alternaria, Cladosporium, Fusarium and Penicillium verrucosum in grain samples harvested in 2011 and 2012 from conventional and organic farms using qPCR. In general, both Alternaria and Cladosporium occurred in all cereal grains in the highest quantities, followed by P. verrucosum and Fusarium. Alternaria, Cladosporium and P. verrucosum had the highest levels in crop mixtures, barley and rye and lower levels in wheat, while Fusarium levels were the highest in crop mixtures and wheat. The levels of Alternaria and P. verrucosum were higher in organic rye and wheat than conventional grains. Although the level of Fusarium was higher in conventional than organic rye, opposite results were obtained for crop mixtures. A positive correlation was found between Alternaria, Cladosporium and P. verrucosum, indicating that similar factors might affect the distribution of these fungi in grains.     

Ovicidal effects of entomopathogenic fungal isolates on the invasive Fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae)

Maize is a major staple food for over 300 million people in sub‐Saharan Africa. Sustainable productivity of this primary crop has been recently threatened by Fall armyworm (FAW), Spodoptera frugiperda invasion. Due to lack of environmentally safe management strategies, immediate responses by growers and governments to tackle FAW are based on rampant use of pesticides. Looking for efficient biopesticides, twenty entomopathogenic fungal isolates (14 Metarhizium anisopliae and 6 Beauveria bassiana) were screened for their efficacy against eggs and second instar larvae of FAW. A single discriminating concentration of 1 × 10⁸ conidia ml^?1 and four replicates per treatment were used in all experiments. Isolates were assessed for their ability to cause mortality of FAW second instar larvae, eggs and the neonate larvae that emerged from treated eggs. Among the isolates tested, only B. bassiana ICIPE 676 caused moderate mortality of 30% to second instar larvae. Metarhizium anisopliae ICIPE 78, ICIPE 40 and ICIPE 20 caused egg mortalities of 87.0%, 83.0% and 79.5%, respectively, and M. anisopliae ICIPE 41 and ICIPE 7 outperformed all the others by causing 96.5% and 93.7% mortality to the neonate larvae, respectively. The cumulated mortality of eggs and neonates was highest with M. anisopliae ICIPE 41 (97.5%), followed by M. anisopliae ICIPE 7, 655, 40, 20 and 78 with total mortality of 96.0%, 95.0%, 93.5%, 93.0% and 92.0%, respectively. These isolates with high cumulated mortality (≥92%), especially ICIPE 78 and 7, which are already commercialized for spider mites and ticks control respectively, would be good candidates for development as biopesticides for management of FAW in Africa if further evidence of their efficacy is obtained in the field.     

Phylogenetic and biogeographic implications inferred by mitochondrial intergenic region analyses and ITS1-5.8S-ITS2 of the entomopathogenic fungi <Emphasis Type="Italic">Beauveria bassiana</Emphasis> and <Emphasis Type="Italic">B. brongniartii</Emphasis>

Background  

The entomopathogenic fungi of the genus Beauveria are cosmopolitan with a variety of different insect hosts. The two most important species, B. bassiana and B. brongniartii, have already been used as biological control agents of pests in agriculture and as models for the study of insect host - pathogen interactions. Mitochondrial (mt) genomes, due to their properties to evolve faster than the nuclear DNA, to contain introns and mobile elements and to exhibit extended polymorphisms, are ideal tools to examine genetic diversity within fungal populations and genetically identify a species or a particular isolate. Moreover, mt intergenic region can provide valuable phylogenetic information to study the biogeography of the fungus.     

The combined effect of soil‐applied azadirachtin with entomopathogens for integrated management of western flower thrips

Performance of soil application of azadirachtin products with entomopathogens was evaluated as single treatments, in multiple combinations and in different concentrations against the soil stages of western flower thrips, Frankliniella occidentalis (Pergande), in French beans Phaseolus vulgaris L. Treatments consisted of NeemAzal‐T solution, neem pellets, Steinernema carpocapsae (Weiser) Nemastar^®, isolates of Metarhizium anisopliae (Metschnikoff) Sorokin (IPP 2539 and ICIPE‐69) and Beauveria bassiana (Balsamo) – Naturalis^®. All treatments were analysed for the number of emerging adults, while emerged adults in fungi‐based treatments were analysed additionally for the retarded development of mycosis as a possible cause of secondary mortality. Possible interactive effects in combined treatments were analysed using a generalized linear model (GLM) approach, and three levels of dose–response combinations of the selected treatments were further tested. Bioassay results of the single treatments indicated between 43% and 60% reduction in adult emergence with NeemAzal‐T solution thus proving to be the most efficient. However, most cadavers with entomopathogenic fungi (EPF) treatments showed the development of mycosis. Therefore, the reduction in adult emergence attributed to the EPF was altogether >87%. Combined treatments with Steinernema, Metarhizium (ICIPE‐69), NeemAzal‐T and neem pellets resulted in total reduction in adult emergence of 95–97% when late mortality by mycosis was considered. Of the treatment combinations, two showed synergistic, four additive and one an antagonistic response. Combining low concentration of entomopathogenic nematode (EPN) (100 IJ/cm²) with NeemAzal‐T resulted in satisfactory control compared to the operational dose of EPN, while the highest concentration of M. anisopliae (10⁸ conidia) combined with Steinernema showed the best performance with 74% reduction in adult emergence.     

Laboratory and field evaluation of entomopathogenic nematodes for control of Agriotes obscurus (L.) (Coleoptera: Elateridae)

The susceptibility of the dusky wireworm, Agriotes obscurus (L.) (Coleoptera: Elateridae), to different species and strains of entomopathogenic nematodes was tested in a virulence assay in the laboratory. Larvae were exposed to different nematode doses of 50 and 100 IJs/cm². At a dose of 50 IJs/cm², only a commercial strain Heterorhabditis bacteriophora Poinar and the native strain Steinernema carpocapsae (Weiser) B14 caused increased mortality compared with the control (11.1% and 13.3% mortality, respectively). At the higher dose tested, all strains (except Steinernema sp. D122) were virulent to A. obscurus larvae. Steinernema carpocapsae B14 caused higher mortality of wireworm (75.6%) and was used for the assay conducted in cages, with a dose of 100 IJs/cm², in field conditions. The results showed that S. carpocapsae B14 controlled 48.3% of A. obscurus larvae, demonstrating that some entomopathogenic nematodes have the potential to control larvae of A. obscurus. However, further work is needed to improve their efficacy.     

Entomopathogenic fungi of Eurygaster integriceps Puton (Hemiptera: Scutelleridae): collection and characterization for development

Exploratory activities were done in Syria, Turkey, Iran, Uzbekistan, Kazakhstan, The Kyrghyz Republic, and Russia to locate entomopathogenic fungi of Eurygaster integriceps. Isolates from the entomopathogenic genera Beauveria, Paecilomyces, and Verticillium were collected. Beauveria bassiana was the most commonly recovered species. Thirty-one isolates of the 221 recovered were examined at 20, 25, 30, and 35 °C for 20 days for growth and sporulation. Growth and sporulation were generally highest at 25 °C. None of the isolates grew at 35 °C, and at 30 °C growth was retarded with no conidia being produced. Single- and multiple-concentration bioassays were conducted on greenhouse-grown wheat plants and in pine litter to evaluate virulence of fungi from several sources to E. integriceps. When tested at a single concentration, mortality after 15 days ranged from 66 to >95% in the litter assays and 50 to 91% in the plant assays. There was a distinct concentration response for most of the isolates tested in the multiple-concentration assay, particularly in the in-litter environment. In litter, mortality tended to develop earlier than in on-plant assays. Several isolates of B. bassiana and one Metarhizium anisopliae displayed consistently high virulence against E. integriceps and were more virulent than two commercial strains. Our results demonstrate the potential of entomopathogenic fungi for management of E. integriceps in overwintering sites and in wheat fields.     

Food web associations and effect of trophic resources and environmental factors on parasitoids expanding their host range into non‐native hosts

Trophic interactions and environmental conditions determine the structure of food webs and the host expansion of parasitoids into novel insect hosts. In this study, we investigate plant–insect–parasitoid food web interactions, specifically the effect of trophic resources and environmental factors on the presence of the parasitoids expanding their host range after the invasion of Chrysodeixis chalcites (Esper) (Lepidoptera: Noctuidae). We also consider potential candidates for biological control of this non‐native pest. A survey of larval stages of Plusiinae (Lepidoptera: Noctuidae) and their larval parasitoids was conducted in field and vegetable greenhouse crops in 2009 and 2010 in various locations of Essex and Chatham‐Kent counties in Ontario, Canada. Twenty‐one plant–host insect–host parasitoid associations were observed among Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae), C. chalcites, and larval parasitoids in three trophic levels of interaction. Chrysodeixis chalcites, an old‐world species that had just arrived in the region, was the most common in our samples. The larval parasitoids Campoletis sonorensis (Cameron) (Hymenoptera: Ichneumonidae), Cotesia vanessae (Reinhard), Cotesia sp., Microplitis alaskensis (Ashmead), and Meteorus rubens (Nees) (all Hymenoptera: Braconidae) expanded their host range into C. chalcites changing the structure of the food web. Copidosoma floridanum (Ashmead) (Hymenoptera: Encyrtidae) was the most common parasitoid of T. ni that was not found in the invasive species. Plant species, host abundance, and agro‐ecosystem were the most common predictors for the presence of the parasitoids expanding their host range into C. chalcites. Our results indicate that C. sonorensis, C. vanessae, and C. floridanum should be evaluated for their potential use in biological control of C. chalcites and T. ni.     

Potential of Metarhizium anisopliae and Beauveria bassiana to control Dinoderus porcellus (Coleoptera: Bostrychidae) infesting yam chips

The beetle Dinoderus porcellus Lesne is a serious storage insect pest that causes important losses by destroying stocks of yam chips. In the aim to found an alternative control method to the use of synthetic insecticides for its management, the virulence of the entomopathogenic fungi Beauveria bassiana (Balsamo) Vuillemin (isolate Bb115) and Metarhizium anisopliae (Metschnikoff) Sorokin (isolate Met 31) against adults of D. porcellus was evaluated under laboratory conditions (25 ± 2 °C and 70 ± 5% RH). Then, the effectiveness of the most virulent entomopathogenic fungus as biological agent against D. porcellus was assessed under farmer storage conditions. For each entomopathogenic fungus isolate, four conidial concentration (0, 10⁵, 10⁷, and 10⁹ conidia/mL) at the dose of 1 µL were inoculated topically on D. porcellus adults (3–5 days old). Observations focused on insect mortality, cadaver sporulation and weight loss of yam chips. Lethal dose and lethal time values were estimated using probit analysis. Both fungal isolates at all conidial dose caused more than 50% mortality on day 7, with the highest mortality (94.44%) achieved using B. bassiana at the 10⁹ conidia/mL. LT₅₀ values for B. bassiana and M. anisopliae isolates were 2.63 and 3.35 days, respectively, while their LT₉₀ values were 6.15 and 9.87 days, respectively. Yielding the lower LD₉₀ values and the highest rates of cadaver sporulation, B. bassiana isolate appeared as the most virulent against D. porcellus. After 3 months of storage, comparatively to the control, the B. bassiana isolate at the highest conidial dose (10⁹ conidia/mL) significantly reduced D. porcellus populations, and weight loss of yam chips. This study revealed the potential of B. bassiana and M. anisoplae isolates as biological control agent against D. porcellus for yam chips protection.     

Diversity of fungal latent pathogens and true endophytes associated with fruit trees in Uruguay

We have explored the fungal diversity in asymptomatic twigs of apple, peach, pear and blueberry trees, with the objective of discerning between true endophytes and latent pathogens. Several fungal genera containing known bark pathogens were found. Seven Diaporthe species—D. oxe, D. infecunda, D. serafiniae, D. phaseolorum, D. terebinthifolii, D. foeniculina and D. brasiliensis—were identified, along with Botryosphaeria dothidea, Neofusicoccum parvum, Neofusicoccum australe, Cytospora sp., Cytospora acaciae and Pestalotiopsis spp. A pathogenicity trial was undertaken to determine the role of these species on apple, pear, blueberry and peach shoots. Diaporthe brasiliensis, D. foeniculina, Diaporthe inconspicua, D. terebinthifolii, Diaporthe sp.1, Cytospora‐like isolates and Pestalotiopsis spp. isolates produced no lesions on inoculated shoots, suggesting that they could be considered true endophytes on their respective hosts. Meanwhile, some of the isolates of Diaporthe—D. oxe, Diaporthe sp.2, D. infecunda and D. serafiniae, B. dothidea, N. parvum and N. australe could be regarded as latent pathogens in their respective hosts as they produced sunken cankers and necrosis on inoculated shoots. These results demonstrate that apple, pear, blueberry and peach healthy shoots can host many known endophytic fungi along with potential wood disease‐causing fungi that should be regarded as latent pathogens.     

Multiple introductions from multiple sources: invasion patterns for an important Eucalyptus leaf pathogen

Many population studies on invasive plant pathogens are undertaken without knowing the center of origin of the pathogen. Most leaf pathogens of Eucalyptus originate in Australia and consequently with indigenous populations available, and it is possible to study the pathways of invasion. Teratosphaeria suttonii is a commonly occurring leaf pathogen of Eucalyptus species, naturally distributed in tropical and subtropical regions of eastern Australia where it is regarded as a minor pathogen infecting older leaves; however, repeated infections, especially in exotic plantations, can result in severe defoliation and tree deaths. Nine polymorphic microsatellite markers were used to assess the genetic structure of 11 populations of T. suttonii of which four where from within its native range in eastern Australia and the remaining seven from exotic Eucalyptus plantations. Indigenous populations exhibited high allele and haplotype diversity, predominantly clonal reproduction, high population differentiation, and low gene flow. The diversity of the invasive populations varied widely, but in general, the younger the plantation industry in a country or region, the lower the diversity of T. suttonii. Historical gene flow was from Australia, and while self‐recruitment was dominant in all populations, there was evidence for contemporary gene flow, with South Africa being the most common source and Uruguay the most common sink population. This points distinctly to human activities underlying long‐distance spread of this pathogen, and it highlights lessons to be learned regarding quarantine.